Linear Polyethylenimine-DNA Nanoconstruct for Corneal Gene Delivery.

Area of Science:

• Biomaterials Science
• Ocular Therapeutics
• Gene Delivery Systems

Background:

• Corneal gene therapy holds potential for treating various eye diseases.
• Efficient and safe gene delivery vectors are crucial for successful ocular gene therapy.
• Polyethylenimine (PEI)-DNA nanoconstructs are being explored for their gene delivery capabilities.

Purpose of the Study:

• To evaluate the efficiency and toxicity of a linear 22-kDa PEI-DNA nanoconstruct for corneal gene delivery.
• To determine the optimal PEI nitrogen-to-DNA phosphate (N:P) ratio for gene transfer.
• To assess the in vitro and in vivo performance and safety of the nanoconstruct.

Main Methods:

• Characterization of PEI-DNA nanoconstructs using gel retardation and zeta potential measurements.
• In vitro assessment of gene transfection efficiency and cytotoxicity (MTT, LDH, ROS assays) in human corneal fibroblasts.
• In vivo gene transfer studies in mouse models, evaluating GFP expression and ocular safety via slit-lamp biomicroscopy.

Main Results:

• The optimal N:P ratio of 30:1 demonstrated the highest gene transfection efficiency with no significant cytotoxicity.
• Substantial green fluorescent protein (GFP) gene delivery into mouse corneas was achieved within 3 days of topical application.
• No adverse ocular effects, including redness, edema, inflammation, or tearing, were observed in treated mice.

Conclusions:

• The 22-kDa linear PEI-DNA nanoconstruct serves as an efficient and well-tolerated vector for corneal gene therapy.
• This nanoconstruct represents a viable platform for developing innovative gene-based nanomedicine for corneal diseases.
• The findings support the potential of PEI-DNA nanoconstructs in advancing ocular gene therapeutic strategies.